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Fig. 4 | Genome Biology

Fig. 4

From: Guide RNAs with embedded barcodes boost CRISPR-pooled screens

Fig. 4

Comparison of the CRISPRiBAR and conventional CRISPR-pooled screens for the identification of human genes important for 6-TG-mediated cytotoxicity in HeLa cells. a, b Screening scores of the top-ranked genes calculated by MAGeCKiBAR (a) and by MAGeCK (b). Identified candidates (FDR < 0.15) were labeled, and only top 10 hits were labeled for MAGeCKiBAR screens. Negative control genes were labeled with black dots. c Validation of reported genes (MLH1, MSH2, MSH6, and PMS2) involved in 6-TG cytotoxicity. d Spearman correlation coefficient of the top 20 positively selected genes between two biological replicates using MAGeCKiBAR (left) or conventional MAGeCK analysis (right). e Validation of top candidate genes isolated by either MAGeCKiBAR or MAGeCK analysis. Mini-pooled sgRNAs targeting each gene were delivered to cells through lentiviral infection. Transduced cells were cultured for an additional 10 days before 6-TG treatment. Data are presented as the mean ± S.E.M. (n = 5). P values were calculated using Student’s t test. *P < 0.05; **P < 0.01; ***P < 0.001; NS, not significant. The sgRNA sequences for validation are listed in Additional file 3: Table. S8. f, g The sgRNAiBAR read counts for HPRT1 targeting (f) and FGF13 targeting (g) before (Ctrl) and after (Exp) 6-TG screening in two replicates

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